Prenatal ultrasound assessment and fetal heart monitoring: analysis of dizygotic twins discordant for body stalk anomaly in the third trimester of pregnancy.
We describe a case of dizygotic twin pregnancy which was referred to our centre because of a discordant anomaly of one of the twins. The ultrasound examination revealed a large extra-abdominal mass (liver and bowel) with micromelia, severe kyphoscoliosis and a short umbilical cord of the second dizygotic discordant twin. These ultrasound findings were diagnosed as body stalk anomaly. Body stalk anomaly in twins is extremely rare. This is, to our knowledge, one of the few documented cases reported on dizygotic twins discordant for this anomaly. This finding, in association with a decrease of amniotic fluid volume, may be attributed to early amniotic membrane rupture as the primary event in the pathogenesis of body stalk anomaly.


A case-control evaluation of pulmonary and extrapulmonary findings of incidental asymptomatic COVID-19 infection on FDG PET-CT.
To describe the findings of incidental asymptomatic COVID-19 infection on FDG PET-CT using a case-control design. Incidental pulmonary findings suspicious of asymptomatic COVID-19 infection on FDG PET-CT were classified as a confirmed (positive RT-PCR test) or suspected case (no/negative RT-PCR test). Control cases were identified using a 4:1 control:case ratio. Pulmonary findings were re-categorised by two reporters using the BSTI classification. SUV metrics in ground glass opacification (GGO)/consolidation (where present), background lung, intrathoracic nodes, liver, spleen and bone marrow were measured. 7/9 confirmed and 11/15 suspected cases (COVID-19 group) were re-categorised as BSTI 1 (classic/probable COVID-19) or BSTI 2 (indeterminate COVID-19); 0/96 control cases were categorised as BSTI 1. Agreement between two reporters using the BSTI classification was almost perfect (weighted κ = 0.94). SUVmax GGO/consolidation (5.1 vs 2.2; p < 0.0001) and target-to-background ratio, normalised to liver SUVmean (2.4 vs 1.0; p < 0.0001) were higher in the BSTI 1 & 2 group vs BSTI 3 (non-COVID-19) cases. SUVmax GGO/consolidation discriminated between the BSTI 1 & 2 group vs BSTI 3 (non-COVID-19) cases with high accuracy (AUC = 0.93). SUV metrics were higher (p < 0.05) in the COVID-19 group vs control cases in the lungs, intrathoracic nodes and spleen. Asymptomatic COVID-19 infection on FDG PET-CT is characterised by bilateral areas of FDG avid (intensity > x2 liver SUVmean) GGO/consolidation and can be identified with high interobserver agreement using the BSTI classification. There is generalised background inflammation within the lungs, intrathoracic nodes and spleen. Incidental asymptomatic COVID-19 infection on FDG PET-CT, characterised by bilateral areas of ground glass opacification and consolidation, can be identified with high reproducibility using the BSTI classification. The intensity of associated FDG uptake (>x2 liver SUVmean) provides high discriminative ability in differentiating such cases from pulmonary findings in a non-COVID-19 pattern. Asymptomatic COVID-19 infection causes a generalised background inflammation within the mid-lower zones of the lungs, hilar and central mediastinal nodal stations, and spleen on FDG PET-CT.


Rapid and cost-effective antenatal diagnosis of haemoglobin Bart's hydrops foetalis syndrome using a duplex-polymerase chain reaction.
Haemoglobin Bart's hydrops foetalis syndrome (--SEA/--SEA) is not compatible with life and contributes to a majority of the hydropic foetuses in the Malaysian Chinese alpha-thalassaemia carriers who possess the 2-alpha-gene deletion in cis (--SEA/alphaalpha). A duplex-PCR which simultaneously amplifies a normal 136 bp sequence between the psialpha-alpha2-globin genes and a 730 bp Southeast Asian deletion-specific sequence (--SEA) between the psialpha2-theta1-globin genes was established. The duplex-PCR which detects the --SEA deletion in both chromosomes serves as a rapid and cost-effective confirmatory test in the antenatal diagnosis of Haemoglobin Bart's hydrops foetalis syndrome in Malaysia. In addition, the duplex-PCR is simple to perform as both the normal and deletion-specific alpha-globin gene sequences are amplified in the same PCR reaction.


Mechanistic Study of Utilization of Water-Insoluble Saccharomyces cerevisiae Glucans by Bifidobacterium breve Strain JCM1192.
Bifidobacteria exert beneficial effects on hosts and are extensively used as probiotics. However, due to the genetic inaccessibility of these bacteria, little is known about their mechanisms of carbohydrate utilization and regulation. Bifidobacterium breve strain JCM1192 can grow on water-insoluble yeast (Saccharomyces cerevisiae) cell wall glucans (YCWG), which were recently considered as potential prebiotics. According to the results of 1H nuclear magnetic resonance (NMR) spectrometry, the YCWG were composed of highly branched (1→3,1→6)-β-glucans and (1→4,1→6)-α-glucans. Although the YCWG were composed of 78.3% β-glucans and 21.7% α-glucans, only α-glucans were consumed by the B. breve strain. The ABC transporter (malEFG1) and pullulanase (aapA) genes were transcriptionally upregulated in the metabolism of insoluble yeast glucans, suggesting their potential involvement in the process. A nonsense mutation identified in the gene encoding an ABC transporter ATP-binding protein (MalK) led to growth failure of an ethyl methanesulfonate-generated mutant with yeast glucans. Coculture of the wild-type strain and the mutant showed that this protein was responsible for the import of yeast glucans or their breakdown products, rather than the export of α-glucan-catabolizing enzymes. Further characterization of the carbohydrate utilization of the mutant and three of its revertants indicated that this mutation was pleiotropic: the mutant could not grow with maltose, glycogen, dextrin, raffinose, cellobiose, melibiose, or turanose. We propose that insoluble yeast α-glucans are hydrolyzed by extracellular pullulanase into maltose and/or maltooligosaccharides, which are then transported into the cell by the ABC transport system composed of MalEFG1 and MalK. The mechanism elucidated here will facilitate the development of B. breve and water-insoluble yeast glucans as novel synbiotics.IMPORTANCE In general, Bifidobacterium strains are genetically intractable. Coupling classic forward genetics with next-generation sequencing, here we identified an ABC transporter ATP-binding protein (MalK) responsible for the import of insoluble yeast glucan breakdown products by B. breve JCM1192. We demonstrated the pleiotropic effects of the ABC transporter ATP-binding protein in maltose/maltooligosaccharide, raffinose, cellobiose, melibiose, and turanose transport. With the addition of transcriptional analysis, we propose that insoluble yeast glucans are broken down by extracellular pullulanase into maltose and/or maltooligosaccharides, which are then transported into the cell by the ABC transport system composed of MalEFG1 and MalK. The mechanism elucidated here will facilitate the development of B. breve and water-insoluble yeast glucans as novel synbiotics.


Clustering irregular shapes using high-order neurons.
This article introduces a method for clustering irregularly shaped data arrangements using high-order neurons. Complex analytical shapes are modeled by replacing the classic synaptic weight of the neuron by high-order tensors in homogeneous coordinates. In the first- and second-order cases, this neuron corresponds to a classic neuron and to an ellipsoidalmetric neuron. We show how high-order shapes can be formulated to follow the maximum-correlation activation principle and permit simple local Hebbian learning. We also demonstrate decomposition of spatial arrangements of data clusters, including very close and partially overlapping clusters, which are difficult to distinguish using classic neurons. Superior results are obtained for the Iris data.


Inhibition of syk activity and degranulation of human mast cells by flavonoids.
To investigate the effect of flavonoids on the activation of p72(syk) (Syk) protein tyrosine kinase which plays a pivotal role in the high affinity IgE receptor-mediated degranulation of mast cells, we picked out 10 flavonoids, classified them into 4 series, and examined their effects on the activation of Syk and on the degranulation of human mast cells. Flavones and flavonols showed clear inhibition, whereas flavanones and isoflavones had either weak or no effect on Syk enzymatic activity induced by amino acid peptide corresponding to the activation loop domain and on IgE-dependent degranulation of human cultured mast cells (HCMC). On the basis of calculated logP (ClogP) values as a prediction of compound lipophilicity, some flavonoids were speculated to have low lipophilicity, the reason for poor cell permeability. A significant relationship was observed between the inhibition of Syk activity and HCMC degranulation attributable to flavonoids when the ClogP values of the compounds were taken into account (r(2)=0.89). These results suggested that the impairment of mast cell degranulation by several flavonoids classified into flavones and flavonols might be mediated via inhibition of the intracellular activation of Syk.


Heterozygous IDH1R132H/WT created by "single base editing" inhibits human astroglial cell growth by downregulating YAP.
Mutations in the isocitrate dehydrogenase 1 (IDH1) gene have been identified in a number of cancer types, including brain cancer. The Cancer Genome Atlas project has revealed that IDH1 mutations occur in 70-80% of grade II and grade III gliomas. Until recently, most of the functional studies of IDH1 mutations in cellular models have been conducted in overexpression systems with the IDH1 wild type background. In this study, we employed a modified CRISPR/Cas9 genome editing technique called "single base editing", and efficiently introduced heterozygous IDH1 R132H mutation (IDH1R132H/WT) in human astroglial cells. Global DNA methylation profiling revealed hypermethylation as well as hypomethylation induced by IDH1R132H/WT. Global gene expression analysis identified molecular targets and pathways altered by IDH1R132H/WT, including cell proliferation, extracellular matrix (ECM), and cell migration. Our phenotype analysis indicated that compared with IDH1 wild type cells, IDH1R132H/WT promoted cell migration by upregulating integrin β4 (ITGB4); and significantly inhibited cell proliferation. Using our mutated IDH1 models generated by "single base editing", we identified novel molecular targets of IDH1R132H/WT, namely Yes-associated protein (YAP) and its downstream signaling pathway Notch, to mediate the cell growth-inhibiting effect of IDH1R132H/WT. In summary, the "single base editing" strategy has successfully created heterozygous IDH1 R132H mutation that recapitulates the naturally occurring IDH1 mutation. Our isogenic cellular systems that differ in a single nucleotide in one allele of the IDH1 gene provide a valuable model for novel discoveries of IDH1R132H/WT-driven biological events.


American Society of Nephrology quiz and questionnaire 2014: glomerular diseases.
The Nephrology Quiz and Questionnaire remains an extremely popular session for attendees of the Annual Kidney Week Meeting of the American Society of Nephrology. Once again, the conference hall was overflowing with audience members and eager quiz participants. Topics covered by the expert discussants included electrolyte and acid-base disorders, glomerular disease, ESRD/dialysis, and transplantation. Complex cases representing each of these categories along with single best answer questions were prepared and submitted by the panel of experts. Before the meeting, program directors of United States nephrology training programs and nephrology fellows answered the questions through an internet-based questionnaire. During the live session, members of the audience tested their knowledge and judgment on a series of case-oriented questions that were prepared and discussed by the experts. They compared their answers in real time using audience response devices with the answers of the nephrology fellows and training program directors. The correct and incorrect answers were then discussed after the audience responses, and the results of the questionnaire were displayed. As always, the audience, lecturers, and moderators enjoyed this educational session. This article recapitulates the session and reproduces its educational value for the readers of CJASN. Enjoy the clinical cases and expert discussions.


Host Plant Associations of Anagrus spp. (Hymenoptera: Mymaridae) and Erythroneura elegantula (Hemiptera: Cicadellidae) in Northern California.
Anagrus erythroneurae S. Trjapitzin & Chiappini and Anagrus daanei Triapitsyn are the key parasitoids of the western grape leafhopper (Erythroneura elegantula Osborn) in northern California vineyards. Erythroneura elegantula overwinters as an adult in reproductive diapause. To successfully overwinter, Anagrus spp. must locate an alternate leafhopper host that overwinters in an egg stage that they can parasitize. These alternate leafhopper hosts are thought to be primarily located in the natural habitats surrounding vineyards. This study identifies the noncrop host plants utilized by Anagrus spp. not only during the overwintering period but throughout the entire year, as well as the leafhopper species associated with these host plants. Over a 2-yr period, Anagrus spp. and leafhoppers were sampled from numerous plants in natural and cultivated habitats surrounding vineyards. Results from this study confirm previously known Anagrus spp. host plants, but also identify new host plant species. Some of the host plants harbored Anagrus spp. year-round while others were utilized only during certain periods of the year. Leafhoppers associated with Anagrus spp. host plants may potentially serve as the alternate host utilized by Anagrus spp. on these plants, but this was not confirmed in the current study. Records of E. elegantula demonstrate their cyclical movement between the vineyard floor (winter), temporary noncrop hosts (spring/fall), and the grape vine canopy (summer).


Meta-analysis of association between ApoE epsilon4 allele and schizophrenia.
Several case-control studies have reported an association between schizophrenia and the epsilon4 allele of Apolipoprotein E gene. The results have been equivocal. This meta-analysis has evaluated the collective evidence for an association between the epsilon4 allele of Apolipoprotein E gene and schizophrenia. We analyzed published data sequentially first considering epsilon4 allele itself, and then epsilon4 carrier status as risk factors for schizophrenia using a sample of 17 population-based case-control studies, of which 6 were from Asian and 11 from Caucasian populations. The pooled odds ratios from the Caucasian populations showed a modest association with risk of schizophrenia for epsilon4 allele and epsilon4 carrier genotype. No other alleles or genotypes were significant in either Asian or Caucasian populations when analysed separately or combined, although the sample size had over 80% power to detect a significant odds ratio of 1.9 in Asian-population studies and 1.6 in Caucasian-population studies. After allowing for sensitivity analysis of the studies and assessment of publication bias, we conclude that the epsilon4 allele of Apolipoprotein E does not play a major role in risk of schizophrenia in Caucasian populations. Since significant heterogeneity was present among the 6 Asian populations reported to date, further studies using larger sample sizes are required.


A qualitative exploration of rural feeding and weaning practices, knowledge and attitudes on nutrition.
An exploratory qualitative investigation was done to determine the feeding and weaning practices, knowledge and attitudes towards nutrition of mothers/caregivers of children up to 3 years old attending baby clinics in the Moretele district (South Africa). Qualitative data collection on six relevant nutrition topics was done using focus group interviews. Trained moderators, using a pre-tested, structured interview schedule, interviewed participants in six age groups. Focus group interviews were taped, transcribed and translated. Content analysis produced systematic data descriptions and ethnography provided descriptive data. Breast-feeding was the choice feed and bottle-feeding was only given when breast-feeding was impossible. Solid food was introduced early (at 2-3 months) and a mixed family diet at 7-9 months. Milk feeds were stopped completely from 18-24 months. Weaning diets were compromised due to poor food choices, preparation practices and limited variety. The participant's nutrition knowledge regarding specific foods, their functions and recommended quantities was poor. The women adhered to their cultural beliefs regarding food choices and preparation practices. The data analysis revealed that inadequate nutrition knowledge and adherence to cultural practices lead to poor-quality feeding practices. Cultural factors and taboos have a powerful influence on feeding practices and eating patterns. Young mothers often find it impossible to ignore their ill-informed elders or peer group. Nutrition knowledge needs to be changed in a first step towards implementing improved feeding practices. Facilitated group discussions could focus on possible solutions for the identified nutrition-related problems.


[REGULATION OF THE mTOR SIGNALING PATHWAY IN MACROPHAGES IN VARIOUS PATHOLOGIES].
Macrophage is a key cell of immune system, it participates in antiviral, antimicrobial and antitumor defense of the organism, also in regeneration and reparation of tissues. Macrophage coordinates functioning of immune system, participates in tumor growth progression. The process of inflammation consists of two stages. Cytotoxical potential of immunocompetent cells will be realized in the first stage, to avoid a bacterial infection. The second stage of inflammatory process is associated with reparation and regeneration. During inflammation, according it stages, macrophages change functional state, switching from cytotoxical M1 to M2, that associated with reparation. We suppose, that rapamysin, a suppressor of mTOR, causes completely different effects on tumor associated macrophages and cells of microglia. Rapamycin transforms tissue macrophages into M1 phenotype, promoting the tumor regression. While in microglial cells of the central nervous system it induces transformation into M2 phenotype, facilitating the course of the neurodegenerative disease and slowing down the aging.


A Vexing Recurrent Pericardial Effusion Without Underlying Pathology - Subsequent Diagnosis Of Lymphoma Proves A Rare First Presentation.
An elderly man had recurrent admissions with large symptomatic pericardial effusions. Initial computed tomography (CT) of thorax, abdomen and pelvis and pericardial fluid analysis did not reveal underlying cause. On subsequent presentation, pericardial window was formed but repeat pericardial fluid analysis and biopsy failed to give a diagnosis again. He then presented approximately after four months with worsening symptoms of dyspnoea and weight loss. General physical examination at that point noted inguinal lymphadenopathy. Repeat imaging with CT and magnetic resonance imaging (MRI) showed features of metastatic malignancy. Tissue diagnosis from inguinal lymph nodes proved to be diffuse large B-cell lymphoma (DLBCL).


Human-modified habitats facilitate forest-dwelling populations of an invasive predator, Vulpes vulpes.
Invasive and over-abundant predators pose a major threat to biodiversity and often benefit from human activities. Effective management requires understanding predator use of human-modified habitats (including resource subsidies and disturbed environments), and individual variation within populations. We investigated selection for human-modified habitats by invasive red foxes, Vulpes vulpes, within two predominantly forested Australian landscapes. We predicted that foxes would select for human-modified habitats in their range locations and fine-scale movements, but that selection would vary between individuals. We GPS-tracked 19 foxes for 17-166 days; ranges covered 33 to >2500 ha. Approximately half the foxes selected for human-modified habitats at the range scale, with some 'commuting' more than five kilometres to farmland or townships at night. Two foxes used burnt forest intensively after a prescribed fire. In their fine-scale nocturnal movements, most foxes selected for human-modified habitats such as reservoirs, forest edges and roads, but there was considerable individual variation. Native fauna in fragmented and disturbed habitats are likely to be exposed to high rates of fox predation, and anthropogenic food resources may subsidise fox populations within the forest interior. Coordinating fox control across land-tenures, targeting specific landscape features, and limiting fox access to anthropogenic resources will be important for biodiversity conservation.


Fifty years of research on self-replication: an overview.
The study of self-replicating structures or machines has been taking place now for almost half a century. My goal in this article is to present an overview of research carried out in the domain of self-replication over the past 50 years, starting from von Neumann's work in the late 1940s and continuing to the most recent research efforts. I shall concentrate on computational models, that is, ones that have been studied from a computer science point of view, be it theoretical or experimental. The systems are divided into four major classes, according to the model on which they are based: cellular automata, computer programs, strings (or strands), or an altogether different approach. With the advent of new materials, such as synthetic molecules and nanomachines, it is quite possible that we shall see this somewhat theoretical domain of study producing practical, real-world applications.


Internet of Things in Pregnancy Care Coordination and Management: A Systematic Review.
The Internet of Things (IoT) has positioned itself globally as a dominant force in the technology sector. IoT, a technology based on interconnected devices, has found applications in various research areas, including healthcare. Embedded devices and wearable technologies powered by IoT have been shown to be effective in patient monitoring and management systems, with a particular focus on pregnant women. This study provides a comprehensive systematic review of the literature on IoT architectures, systems, models and devices used to monitor and manage complications during pregnancy, postpartum and neonatal care. The study identifies emerging research trends and highlights existing research challenges and gaps, offering insights to improve the well-being of pregnant women at a critical moment in their lives. The literature review and discussions presented here serve as valuable resources for stakeholders in this field and pave the way for new and effective paradigms. Additionally, we outline a future research scope discussion for the benefit of researchers and healthcare professionals.


Aberrant expression of HOX genes in human invasive breast carcinoma.
HOX genes are known not only as master genes that control the morphogenesis, but also as regulator genes that maintain tissue or organ specificity in the adult body. We hypothesized that dysregulated expression of HOX genes was associated with tumor development and malignant progression such as invasion and metastasis. In this study, we analyzed the expression patterns of 39 HOX genes in human invasive ductal breast cancer tissues and normal tissues by the real-time RT-PCR method. We found 11 HOX genes (HOXA1, A2, A3, A5, A9, C11, D3, D4, D8, D9 and D10) expression levels of which were significantly different between cancerous and normal tissues. All 10 genes except HOXC11 were expressed at lower levels in cancerous tissues than normal tissues. Comparing expression levels of each HOX gene among the different types of cancer tissues, the expression level of HOXB7 was lower in lymph node metastasis-positive cancer tissues than negative cancer tissues; those of HOXD12 and D13 were higher in progesterone receptor-positive cancer tissues than negative cancer tissues; and the expression level of HOXC5 was lower in cancerous tissues with mutated-type p53 than in normal and cancerous tissues with wild-type p53. These results suggest that the aberrant expression of HOX genes is related to the development of breast cancer and malignant behavior of cancer cells.


Topical acetazolamide and methazolamide delivered by contact lenses.
Topical acetazolamide has been previously found to be ineffective in lowering intraocular pressure (IOP). Using high-water-content soft contact lenses (Sauflon PW) soaked in acetazolamide, we observed a statistically significant ipsilateral decrease in IOP of 6.3 +/- 0.4 mm Hg in the treated eyes of albino rabbits. The duration of the effect was up to 7 1/2 hours. Methazolamide-soaked contact lenses produced a maximum unilateral reduction of similar magnitude but shorter duration. Both serum and aqueous humor analyses for pH, carbon dioxide pressure, bicarbonate, and base excess indicate that acetazolamide delivered by soft contact lenses is able to penetrate the cornea in sufficient concentration to lower IOP by a local mechanism in rabbits without significant systemic absorption.


Control of hair growth using long-pulsed alexandrite laser is an efficient and cost effective therapy for patients suffering from recurrent pilonidal disease.
Pilonidal sinus (PNS) and its surgical management have a profound impact on hospital resources in terms of finances and productive man-hours. Surgical treatment has been the mainstay of treatment of both acute and chronic pilonidal sinus but recurrence is common. The control of hair growth in the sinus region plays an important role in preventing recurrence. Here, we discuss our experience of treating 19 patients suffering from recurrent pilonidal sinus with laser depilation and its long-term cost effectiveness. This is a retrospective study on patients who had recurrence of pilonidal sinus following multiple surgical treatments. They were treated using long-pulsed alexandrite laser for depilation in the sinus area, an outpatient procedure. Their clinical characteristics and outcomes were then evaluated. There was a significant reduction in hair density after laser treatment (p < 0.001). The disease-free period after laser treatment was significantly longer than that one after surgical treatment (p < 0.001). The average cost of repeated surgical treatment per disease-free month was significantly higher than that of laser treatment (p < 0.001). Evidence suggests the role of natal cleft hair growth in the evolution of the pilonidal disease; therefore, control of hair growth should be considered as an adjunct to the initial treatment via surgery. Compared to surgical treatment of recurrences, laser depilation is an efficient and cost-effective method of preventing recurrence and reducing morbidity and loss of man-hours. We suggest that laser depilation of the pilonidal sinus should be funded by clinical commissioning groups.


Molecular detection of P. vivax and P. ovale foci of infection in asymptomatic and symptomatic children in Northern Namibia.
Knowledge of the foci of Plasmodium species infections is critical for a country with an elimination agenda. Namibia is targeting malaria elimination by 2020. To support decision making regarding targeted intervention, we examined for the first time, the foci of Plasmodium species infections and regional prevalence in northern Namibia, using nested and quantitative polymerase chain reaction (PCR) methods. We used cross-sectional multi-staged sampling to select 952 children below 9 years old from schools and clinics in seven districts in northern Namibia, to assess the presence of Plasmodium species. The median participant age was 6 years (25-75%ile 4-8 y). Participants had a median hemoglobin of 12.0 g/dL (25-75%ile 11.1-12.7 g/dL), although 21% of the cohort was anemic, with anemia being severer in the younger population (p<0.002). Most of children with Plasmodium infection were asymptomatic (63.4%), presenting a challenge for elimination. The respective parasite prevalence for Plasmodium falciparum (Pf), Plasmodium vivax (Pv) and Plasmodium ovale curtisi (Po) were (4.41%, 0.84% and 0.31%); with Kavango East and West (10.4%, 6.19%) and Ohangwena (4.5%) having the most prevalence. Pv was localized in Ohangwena, Omusati and Oshana, while Po was found in Kavango. All children with Pv/Pf coinfections in Ohangwena, had previously visited Angola, affirming that perennial migrations are risks for importation of Plasmodium species. The mean hemoglobin was lower in those with Plasmodium infection compared to those without (0.96 g/dL less, 95%CI 0.40-1.52 g/dL less, p = 0.0009) indicating that quasi-endemicity exists in the low transmission setting. We conclude that Pv and Po species are present in northern Namibia. Additionally, the higher number of asymptomatic infections present challenges to the efforts at elimination for the country. Careful planning, coordination with neighboring Angola and execution of targeted active intervention, will be required for a successful elimination agenda.


Multiple preprosomatostatin sorting signals mediate secretion via discrete cAMP- and tetradecanoylphorbolacetate-responsive pathways.
We have previously detected a sorting signal in the amino-terminal 78 residues of rat preprosomatostatin (rPPSS) that targets the precursor into a regulated secretory pathway or pathways allowing proteolytic maturation (Sevarino, K. A., Stork, P., Ventimiglia, R., Mandel, G., and Goodman, R. H. (1989) Cell 57, 11-19). To further localize this signal, we constructed three rPPSS expression vectors that code for substitutions or mutations spanning that portion of rPPSS implicated in sorting, and the precursors were expressed in RIN 5F cells. Fractionation of the intracellular products revealed that accurate processing to somatostatin-14 (SS-14) was not affected by any of the mutations. Examination of the secreted products showed no reduction in processing efficiency, indicating that none of the mutations blocked sorting from constitutive into regulated secretion. Finally, we examined the response to two separate secretogogues, cAMP and 12-O-tetradecanoylphorbol-13-acetate (TPA). Clones expressing two of the three mutant precursors displayed the same stimulation of SS-14 secretion by exogenously administered cAMP and TPA as cells expressing wild-type rPPSS, indicating that targeting specifically to the secretory pathway, or pathways, responsive to cAMP and TPA was not disrupted. However, cells expressing the mutant precursor containing a substitution of the amino-terminal 34 residues of rPPSS by the amino terminus of the vesicular stomatitis virus G protein displayed greatly reduced stimulation of SS-14 secretion by TPA, with a less than compensatory increase in response to cAMP, when compared to cells expressing wild-type rPPSS. In conjunction with our previous studies with anglerfish preprosomatostatins, we conclude that 1) the sorting signal(s) in rPPSS necessary for cAMP-responsive secretion are redundant and probably reside within both mature peptide regions and extrapeptide regions; 2) two or more distinct regulated secretory pathways utilized by secreted peptides can be demonstrated in transfected endocrine cells and targeting to these pathways can be separately mediated by at least two different types of sorting signals within the neuropeptide precursor itself; and 3) pro-region conformation plays little role in prosomatostatin-processing site recognition.


The influence of diluted seawater and ripening stage on the content of antioxidants in fruits of different tomato genotypes.
The aim of this study was to investigate if the combined effect of diluted seawater and ripening can improve the beneficial nutritional properties of tomato fruits from an antioxidant point of view. To reach the goal, different tomato cultivars and breeding lines, genetically modified for ripening, were investigated, and analysis of NADPH and NADP+ as well as of the main antioxidants such as ascorbic acid, lipoic acid, and tocopherols was performed at two ripening stages. The research was conducted on berries of the following genotypes of tomato: cv. Jama, Gimar wild type, Gimar gf, and Gimar nor. The mutant gf is a typical "stay green" mutant, characterized by an incomplete loss of chlorophyll; the nor mutation is characterized by a reduced biosynthesis of ethylene and carotenoids. Both ripening and salinity induced an oxidative stress, and the sensitivity to salt treatment was genotype-dependent. The genotypes cv. Jama and Gimar gf line showed increases in ascorbic acid, lipoic acid, and alpha-tocopherol during both ripening and salt treatment whereas total ascorbate and tocopherols decreased in the berries from salt-treated plants of Gimar wild type. Ripening also determined decreases in ascorbate and tocopherol amounts in the Gimar nor line where a positive effect of ripening and salinity was observed.


FREEZING-OUT TECHNIQUE APPLIED TO THE CONCENTRATION OF BIOLOGICALLY ACTIVE MATERIALS.
When applied to a dilute solution of folic acid and glucose, a freezing-out (with agitation) technique was shown to be an effective method of achieving a 20-fold reduction in volume with a loss of 10% of the active material being concentrated. Concentration of a stimulatory factor for Lactobacillus casei produced by Candida albicans in a complex medium was limited by the total solute concentration. Salts in the medium were concentrated to levels inhibitory for L. casei. The process is not selective and all solutes are concentrated.


The role of WDR76 protein in human diseases.
The WD40 repeat (WDR) domain is one of the most abundant protein interaction domains in the human proteome. More than 360 protein interaction domains have been annotated thus far. The WDR domains mediate interactions with peptide regions of important interaction partners in a variety of biological processes. Proteins with the WDR domain which typically contains a seven-bladed β propeller, are continuously being discovered. They represent a large class of proteins that are likely to play important roles. WD40 repeat domain-containing protein 76 (WDR76) is a member of WDR domain-containing proteins. Although it remains poorly understood, it is potentially involved in DNA damage repair, apoptosis, cell cycle progression, and gene expression regulation. Ongoing research on WDR76 is increasing the knowledge regarding its basic functions and role in different pathophysiological. The study of WDR76 is challenging due to the complexity of its interactions with its partners. In the present review, we summarized the current knowledge regarding WDR76, its physiological functions, the close relationship with human diseases, and potential opportunities for target therapy.


Treg/Th17 polarization by distinct subsets of breast cancer cells is dictated by the interaction with mesenchymal stem cells.
Breast cancer (BC) cells (BCCs) exist within a hierarchy beginning with cancer stem cells (CSCs). Unsorted BCCs interact with mesenchymal stem cells (MSCs) to induce regulatory T cells (Tregs). This study investigated how distinct BCC subsets interacted with MSCs to polarize T-cell response, Tregs versus T helper 17 (Th17). This study tested BC initiating cells (CSCs) and the relatively more mature early and late BC progenitors. CSCs interacted with the highest avidity to MSCs. This interaction required CXCR4 and connexin 43 (Cx43)-dependant gap junctional intercellular communication (GJIC). This interaction induced Treg whereas interactions between MSCs and the progenitors induced Th17 response. The increases in Treg and Th17 depended on MSCs but not CTLA-4, which was increased in the presence of MSCs. Studies with BM stroma (fibroblasts) and MSCs from the same donors, indicated specific effects of MSCs. In total, MSC-CSC interaction required CXCR4 for GJIC. This led to increased Tregs and TGFβ, and decreased Th17. In contrast, late and early BCCs showed reduced formation of GJIC, decreased Treg and increased Th17 and IL-17. These findings have significance to the methods by which CSCs evade the immune response. The findings could provide methods of intervention to reverse immune-mediated protection and support of BC.


Concomitant pulmonologist and primary care for chronic obstructive pulmonary disease: a population study.
Pulmonologists provide quality care, however, their number is not adequate to take care of all the chronic obstructive pulmonary disease (COPD) needs of the population and their services come with a cost. Their optimal role should be defined, ideally based on evidence, to ensure that their abilities are applied most efficiently where needed. To determine if concomitant pulmonologist and primary care physician care after COPD hospital or emergency department discharge was associated with better health outcomes than primary care services alone. A population cohort study was conducted in Ontario, Canada from 2004 to 2011. All individuals with a COPD hospital or emergency department discharge were included. Patients who visited both a pulmonologist and a primary care physician within 30 days of the index discharge were matched to patients who had visited a primary care physician alone using propensity scores. The composite outcome of death, COPD hospitalization or COPD emergency department visit was compared using proportional hazards regression. In the propensity score matched sample, 39.7% of patients who received concomitant care and 38.9% who received primary care only died or visited the emergency department visit or hospital for COPD within 1 year (adjusted hazard ratio 1.08, 95% confidence interval 1.00-1.17). The former, however, were more likely to receive diagnostic testing and medications. Patients who received concomitant care after COPD emergency department or hospital discharge did not have better outcomes than those who received primary care alone, however, they did receive more testing and medical management.


Stereotactic body radiotherapy for central lung tumors.
Patients with centrally located lung tumors have been reported to have a higher risk of toxicity when treated with stereotactic body radiotherapy (SBRT) compared with patients with peripheral tumors. The optimal SBRT fractionation schedule for treatment of central tumors is unknown. The primary purpose of this study was to assess toxicity in patients with central lesions treated with SBRT at our institution, the majority of whom were treated with four fractions. Forty-seven patients with 51 central lesions, either primary lung cancer or lung metastases, were treated with SBRT at the Department of Therapeutic Radiology, Yale University School of Medicine/Yale Cancer Center from 2007 to 2011. The patients were treated with three to five fractions with the majority of patients receiving 50 Gy in four fractions of 12.5 Gy. Forty of the lesions were located within 2 cm of the proximal tracheobronchial tree whereas 11 were located within 2 cm of other mediastinal structures. Toxicity data were collected and analyzed according to pretreatment and tumor characteristics and dosimetric parameters. Lobar control data were compiled. With a median follow-up of 11.3 months (range, 4.8-40.8), four patients experienced grade 3 dyspnea and one patient developed hemoptysis that contributed to respiratory failure and subsequent death. Grade 2 toxicity included fatigue (n = 3), dyspnea (n = 3), chest-wall pain (n = 1), and cough (n = 1). Patients with grade 3+ toxicity had larger maximum tumor diameters compared with those patients without grade 3+ toxicity (median diameter 4.3 cm versus 2.9 cm, p = 0.02). There were no detectable significant differences between the two groups with respect to baseline pulmonary function tests, distance to tracheobronchial tree, maximum point dose to the tracheobronchial tree, maximum dose to 5 cc of the tracheobronchial tree, mean lung dose, and volume of lung receiving 5 Gy, 10 Gy, and 20 Gy. There were two patients who experienced local recurrences. The median biological equivalent dose (linear quadratic formula, α/β = 10) for patients with local recurrence was 76 Gy compared with 112.5 Gy for patients without local recurrence (2-tailed t test, p = 0.04). The 2-year actuarial lobar local control for the entire cohort was 94%. The 2-year lobar local-control rate for patients receiving a biological equivalent dose of 100 Gy or more was 100% and for those receiving less than 100 Gy was 80% (log rank, p = 0.02). SBRT for central lung tumors seems to be safe, although treatment of larger tumors does carry an increased risk of high-grade toxicity. Efforts to decrease the toxicity risk by decreasing the biologically equivalent dose resulted in increased local failure.


Resolution of ACE inhibitor cough: changes in subjective cough and responses to inhaled capsaicin, intradermal bradykinin and substance-P.
1. In eight hypertensive patients with ACE inhibitor-induced cough the resolution of the cough was examined in a prospective observational study over 4 weeks duration. Resolution of cough was measured by visual analogue scales and questionnaire at baseline and days 3, 7, 14 and 28. In addition changes in cough sensitivity to inhaled capsaicin, and skin responses to bradykinin and substance-P were measured at the same time points. 2. All patients recorded significant subjective improvement in cough questionnaire scores for severity and night time waking, and by visual analogue scales for severity and frequency of cough (all P < 0.0005 for trend from day 0-28). Significant changes in subjective measures were recorded by 3 to 7 days for most measures, but further reductions were observed up to day 28 (all P < 0.01 day 28 vs day 0). 3. The sensitivity to inhaled capsaicin fell over the 28 days of study after stopping enalapril. The potency of capsaicin relative to day 0 was reduced to 0.25 (95% CI 0.07-0.87) by day 14, and to 0.20 (95% CI 0.06-0.67) by 28 days. 4. After stopping enalapril there was a highly significant reduction in wheal area produced by intradermal bradykinin, with the majority of the effect seen by day 3 (P < 0.0005). The wheal area to intradermal substance-P also declined with time after stopping enalapril, but significant changes were not observed until 14 days (P < 0.01). 5. Multiple regression analysis of the rates of decline for the subjective and objective measures of cough showed significant associations between the response to inhaled capsaicin and the VAS scores for severity of cough (P = 0.005) and frequency of cough (P = 0.011). Capsaicin response was not related significantly to the severity of cough measured by self-administered questionnaire. 6. There was a significant association between bradykinin response and VAS scores for frequency of cough (P < 0.04) and severity of cough (P < 0.05), but not with cough by questionnaire or the capsaicin response. The response to substance-P did not relate significantly to any of the measures of cough. 7. Cough caused by enalapril improved markedly by 14 days but took up to 28 days to resolve. It was associated with increased sensitivity to inhaled capsaicin which decreased over 28 days, and which paralleled changes in subjective cough scores.


Cilostazol ameliorates nephropathy in type 1 diabetic rats involving improvement in oxidative stress and regulation of TGF-Beta and NF-kappaB.
Diabetic nephropathy is characterized as the progressive development of renal insufficiency in a setting of hyperglycemia. Previous studies indicate that reactive oxygen species (ROS) play an important role in high glucose-induced renal injury. Cilostazol was reported to lower the production of superoxide significantly in situ. We hypothesized that cilostazol administration in streptozotocin-induced diabetic rats exerts effects via improving oxidative stress. Male Sprague-Dawley rats were fed with cilostazol (5 mg/kg or 25 mg/kg) for 12 weeks after streptozotocin-induced diabetes mellitus. The results showed that cilostazol decreased reactive oxygen species activity significantly in the kidneys of diabetic rats and improved the urine albumin/creatinine ratio. Cilostazol can also improve the levels of serum cholesterol, triglyceride, and LDL-cholesterol. Additionally, diabetes-caused increased glomerular size, TGF-beta, and NF-kappaB decreased under treatment with cilostazol in diabetic rats. Our results indicate that cilostazol has beneficial effects in early diabetic nephropathy.


Long-term activity recording in small aquatic animals.
We have built an activity-recording device based on the principle of detecting slight changes in the heat conductance of an aquatic environment by means of a thermistor. A major advantage of the device is that it can be used with a simple event recorder, thus combining high sensitivity with low cost. The equipment has been tested with automatic mechanical stimulation, and has been used for long-term recording of the swimming of small marine crustaceans. Problems of stability and reliability have been solved, and potential applications of the device seem widespread.


Catalyst design insights from modelling a titanium-catalyzed multicomponent reaction.
High oxidation state transition metal catalysis touches our daily lives through bulk chemical production, e.g. olefin polymerization, and through specialty chemical reactions common in organic synthesis, e.g. the Sharpless asymmetric epoxidation and olefin dihydroxylation. Our group has been expanding the reaction chemistry of titanium(iv) to produce a host of nitrogen-based heterocycles via multicomponent coupling reactions. One such multicomponent coupling reaction discovered in our laboratory is iminoamination, involving an amine, an alkyne, and an isonitrile. However, the experimental modeling of high oxidation state reactions lags far behind that of low oxidation state systems, where a great deal is known about ligands, their donor properties and how their structures affect catalysis. As a result, we have developed an experimental method for determining the donor abilities of anionic ligands on high oxidation state systems, which is based on the chromium(vi) nitride system NCr(NiPr2)2X, where X = the ligand being interrogated. The parameters obtained are simply called ligand donor parameters (LDP). In this contribution, a detailed optimization of the Ti(NMe2)2(dpm)-catalyzed iminoamination reaction was carried out, where dpm = 5,5-dimethyldipyrrolylmethane. During the course of these studies, dimeric {Ti(μ-N-tolyl)(dpm)}2 was isolated, which is proposed as the resting state of the catalyst. To destabilize this resting state, a more electron-rich bis(aryloxide) catalyst system was investigated. The more electron-rich system is somewhat more active for iminoamination under some conditions; however, the catalyst is prone to disproportionation. A study of heteroleptic titanium complexes revealed that the disproportionation equilibrium constant can be effectively modeled as a function of the square of the difference in LDP between the ligands, (ΔLDP)2. Using this methodology, one can estimate the stability of titanium complexes toward disproportionation.


Uptake of intermittent preventive treatment and pregnancy outcomes: health facilities and community surveys in Chókwè district, southern Mozambique.
Malaria in pregnancy leads to serious adverse effects on the mother and the child and accounts for 75,000-200,000 infant deaths every year. Currently, the World Health Organization recommends intermittent preventive treatment of malaria in pregnancy (IPTp) with sulfadoxine-pyrimethamine (SP) at each scheduled antenatal care (ANC) visit. This study aimed to assess IPTp-SP coverage in mothers delivering in health facilities and at the community. In addition, factors associated with low IPTp-SP uptake and malaria adverse outcomes in pregnancy were investigated. A community and a health facility-based surveys were conducted in mothers delivering in Chókwè district, southern Mozambique. Social-demographic data, malaria prevention practices and obstetric history were recorded through self-report and antenatal records. For women delivering at health facilities, a clinical examination of mother and child was performed, and malaria infection at delivery was determined by rapid diagnostic test, microscopy, quantitative PCR and placental histology. Of 1141 participants, 46.6, 30.2, 13.5 and 9.6% reported taking ≥ 3, two, one and none SP doses, respectively. Low IPTp uptake (< 3 doses) was associated with non-institutional deliveries (AOR = 2.9, P < 0.001), first ANC visit after week 28 (AOR = 5.4, P < 0.001), low awareness of IPTp-SP (AOR = 1.6, P < 0.002) and having no or only primary education (AOR = 1.3, P = 0.041). The overall prevalence of maternal malaria (peripheral and/or placental) was 16.8% and was higher among women from rural areas compared to those from urban areas (AOR = 1.9, P < 0.001). Younger age (< 20 years; AOR = 1.6, P = 0.042) and living in rural areas (AOR = 1.9, P < 0.001) were predictors of maternal malaria at delivery. Being primigravidae (AOR = 2.2, P = 0.023) and preterm delivery (AOR = 2.6, P < 0.001) predicted low birth weight while younger age was also associated with premature delivery (AOR = 1.4, P = 0.031). The coverage for two and ≥ 3 doses of IPTp-SP is moderately higher than estimates from routine health facility records in Gaza province in 2015. However, this is still far below the national target of 80% for ≥ 3 doses. Ongoing campaigns aiming to increase the use of malaria prevention strategies during pregnancy should particularly target rural populations, increasing IPTp-SP knowledge, stimulate early visits to ANC, improve access to health services and the quality of the service provided.


Does Age-Related Macular Degeneration (AMD) Treatment Influence Patient Falls and Mobility? A Systematic Review.
Age-related macular degeneration (AMD), a leading cause of irreversible blindness, increases fall risk through impaired central vision. Falls place an enormous economic burden on healthcare systems. We hypothesized that AMD treatments may reduce patients' falls risk. This systematic review (ID #: 172623) synthesized the current understanding of wet and dry AMD treatments' impact on patient falls and mobility, connecting these two public health issues. On April 17, 2020, PubMed, Scopus, CINAHL, and the Cochrane Central Register of Controlled Trials were queried. Clinical trials and observational studies were included, while non-English and non-primary studies were excluded. Two authors screened, extracted data, and assessed bias using RoB-2 and ROBINS-I. A third author served as a tie breaker. This database search resulted in 3,525 studies, with an additional 112 identified through bibliography review. Ten articles met eligibility criteria. Most studies featured the outcome of interest as a secondary outcome (n = 4) and patient-reported adverse events (n = 5), rather than a primary focus (n = 2). Ten out of the 11 outcomes had a moderate to serious risk of bias. No two studies used the same instrument to measure falls or mobility. Despite the potential positive impact of AMD treatments on patient falls and mobility, quality data on this relationship are lacking. This work underscores the need to broaden ophthalmologic research outcomes beyond visual parameters to include patient-centred, functional measures. Incorporating standardized methods to track falls and screen for difficulty with walking and balance would enable evaluation of AMD treatments on functional outcomes, potentially helping guide management.


Dopamine excites fast-spiking interneurons in the striatum.
The striatum is the main recipient of dopaminergic innervation. Striatal projection neurons are controlled by cholinergic and GABAergic interneurons. The effects of dopamine on projection neurons and cholinergic interneurons have been described. Its action on GABAergic interneurons, however, is still unknown. We studied the effects of dopamine on fast-spiking (FS) GABAergic interneurons in vitro, with intracellular recordings. Bath application of dopamine elicited a depolarization accompanied by an increase in membrane input resistance (an effect that persisted in the presence of tetrodotoxin) and action-potential discharge. These effects were mimicked by the D1-like dopamine receptor agonist SKF38393 but not by the D2-like agonist quinpirole. Evoked corticostriatal glutamatergic synaptic currents were not affected by dopamine. Conversely, GABAergic currents evoked by intrastriatal stimulation were reversibly depressed by dopamine and D2-like, but not D1-like, agonists. Cocaine elicited effects similar to those of dopamine on membrane potential and synaptic currents. These results show that endogenous dopamine exerts a dual excitatory action on FS interneurons, by directly depolarizing them (through D1-like receptors) and by reducing their synaptic inhibition (through presynaptic D2-like receptors).


The function of mitogen-activated protein kinase phosphatase-1 in peptidoglycan-stimulated macrophages.
Mitogen-activated protein (MAP) kinases play a pivotal role in the macrophages in the production of proinflammatory cytokines triggered by lipopolysaccharides. However, their function in the responses of macrophages to Gram-positive bacteria is poorly understood. Even less is known about the attenuation of MAP kinase signaling in macrophages exposed to Gram-positive bacteria. In the present study, we have investigated the regulation of MAP kinases and the role of MAP kinase phosphatase (MKP)-1 in the production of pro-inflammatory cytokines using murine RAW264.7 and primary peritoneal macrophages after peptidoglycan stimulation. Treatment of macrophages with peptidoglycan resulted in a transient activation of JNK, p38, and extracellular signal-regulated kinase. Most interestingly, MKP-1 expression was potently induced by peptidoglycan, and this induction was concurrent with MAP kinase dephosphorylation. Triptolide, a diterpenoid triepoxide, potently blocked the induction of MKP-1 by peptidoglycan and prolonged the activation of JNK and p38. Overexpression of MKP-1 substantially attenuated the production of tumor necrosis factor (TNF)-alpha induced by peptidoglycan, whereas knockdown of MKP-1 by small interfering RNA substantially increased the production of both TNF-alpha and interleukin-1 beta. Finally, we found that in primary murine peritoneal macrophages, MKP-1 induction following peptidoglycan stimulation also coincided with inactivation of JNK and p38. Blockade of MKP-1 induction resulted in a sustained activation of both JNK and p38 in primary macrophages. Our results reveal that MKP-1 critically regulates the expression of TNF-alpha and interleukin-1 beta in RAW264.7 cells and further suggest a central role for this phosphatase in controlling the inflammatory responses of primary macrophages to Gram-positive bacterial infection.


A uridylate tract mediates efficient heterogeneous nuclear ribonucleoprotein C protein-RNA cross-linking and functionally substitutes for the downstream element of the polyadenylation signal.
Every RNA added to an in vitro polyadenylation extract became stably associated with both the heterogeneous nuclear ribonucleoprotein (hnRNP) A and C proteins, as assayed by immunoprecipitation analysis using specific monoclonal antibodies. UV-cross-linking analysis, however, which assays the specific spatial relationship of certain amino acids and RNA bases, indicated that the hnRNP C proteins, but not the A proteins, were associated with downstream sequences of the simian virus 40 late polyadenylation signal in a sequence-mediated manner. A tract of five consecutive uridylate residues was required for this interaction. The insertion of a five-base U tract into a pGEM4 polylinker-derived transcript was sufficient to direct sequence-specific cross-linking of the C proteins to RNA. Finally, the five-base uridylate tract restored efficient in vitro processing to several independent poly(A) signals in which it substituted for downstream element sequences. The role of the downstream element in polyadenylation efficiency, therefore, may be mediated by sequence-directed alignment or phasing of an hnRNP complex.


The beginning of the end of the lung cancer epidemic in Dutch women?
In some European countries, female lung cancer mortality and incidence have started to decrease or flatten out, whereas they are still rising in The Netherlands. We present recent mortality and incidence trends of lung cancer and smoking trends in The Netherlands to show the end of the lung cancer epidemic in Dutch women. Lung cancer mortality and incidence rates by gender were analyzed for 4 age groups (20-44, 45-49, 50-54 and 55-59), and smoking prevalence rates were examined for women using joinpoint regression and birth cohort analysis. Data on mortality were collected for the period 1960-2006, incidence for the period 1989-2003 and smoking prevalence for the period 1988-2007. Because of decreasing lung cancer mortality and incidence rates among males and dramatically increasing rates among females, rates of young males were surpassed by those of females after the mid-1990s. However, although in young women (20-49) mortality increased with 4-5% per year, it flattened out (no significant in- or decreases) since 1999. Among older women, mortality rates were still increasing markedly. Mortality rates and smoking prevalence tended to decrease in women born after the 1950s. This is the first report suggesting that the lung cancer epidemic in Dutch women is coming to an end. Although the increase in lung cancer incidence and mortality among Dutch women has been one of the most dramatic in Europe, the recent decrease in young women is expected to be followed by a future leveling off or a slight decrease in overall female lung cancer rates.


[Helicobacter pylori infection and state of antioxidant protection in patients with the unstable course of ischemic disease].
92 patients with ischemic heart disease (IHD) have been examined. Unlike stable angina pectoris, unstable angina and acute myocardial infarction occur more often against the aggravation of chronic atrophic gastritis with a high degree of H. pylori colonization in the mucous coat of the antral part of the stomach. The development of repeated coronary phenomena within one year in patients with unstable angina and acute myocardial infarction was related to the high degree of H. pylori colonization in the mucous coat of the stomach. Irrespective of the presence of IHD in patients, increased H. pylori colonization in the mucous coat was accompanied by a drop in the antioxidant protection activity against the growth of the blood malonic dialdehyde content.


Ultraviolet photodissociation for characterization of whole proteins on a chromatographic time scale.
Intact protein characterization using mass spectrometry thus far has been achieved at the cost of throughput. Presented here is the application of 193 nm ultraviolet photodissociation (UVPD) for top down identification and characterization of proteins in complex mixtures in an online fashion. Liquid chromatographic separation at the intact protein level coupled with fast UVPD and high-resolution detection resulted in confident identification of 46 unique sequences compared to 44 using HCD from prepared Escherichia coli ribosomes. Importantly, nearly all proteins identified in both the UVPD and optimized HCD analyses demonstrated a substantial increase in confidence in identification (as defined by an average decrease in E value of ∼40 orders of magnitude) due to the higher number of matched fragment ions. Also shown is the potential for high-throughput characterization of intact proteins via liquid chromatography (LC)-UVPD-MS of molecular weight-based fractions of a Saccharomyces cerevisiae lysate. In total, protein products from 215 genes were identified and found in 292 distinct proteoforms, 168 of which contained some type of post-translational modification.


Surgical treatment of cervical cord compression in rheumatoid arthritis.
Cervical myelopathy is a rare but potentially dangerous complication of rheumatoid arthritis and presents considerable therapeutic problems. A conservative approach carries high mortality and surgical intervention is not without serious risks. Reduction of subluxation and posterior fusion is widely practised but may require prolonged bed rest and continuous skull traction, sometimes for many weeks. When anterior decompression has been attempted prolonged immobilisation and external fixation have created problems. In this series 23 rheumatoid patients with cervical myelopathy were investigated over a four-year period. Seventeen underwent anterior decompression of the cervical cord, of whom 14 had a transoral removal of the odontoid peg and pannus and posterior occipitocervical fusion during the same anaesthetic without mortality or serious postoperative complications; all but one have improved. The authors believe that early mobilisation after a combined cord decompression and internal fixation has reduced the mortality and morbidity. Management of cervical myelopathy in rheumatoid arthritis and indications for operation are discussed.


Effects of serotonin on the internal anal sphincter: in vivo manometric study in rats.
The effects of serotonin (5-hydroxytryptamine, 5-HT) on the internal anal sphincter were studied in anaesthesized rats. Serotonin induced a dose dependent relaxation of the internal anal sphincter. Methysergide blocked this relaxation, but did not affect the rectoanal reflex. Methysergide did not antagonise the actions of cholinergic and adrenergic agonists on the internal anal sphincter. Other 5-HT antagonists such as cyproheptadine, ketanserin, chlorpromazine, amitriptyline and ergotamine failed to reduce or block the internal anal sphincter relaxation due to 5-HT, nor did they alter the rectoanal reflex. Adrenergic and cholinergic antagonists had no effect on the 5-HT-induced relaxation of the internal anal sphincter, suggesting that 5-HT acts upon the internal anal sphincter via a non-adrenergic, non-cholinergic mechanism. Tetrodotoxin reduced or blocked the relaxation of the internal anal sphincter produced by 5-HT, implying that 5-HT acts through neural pathways rather than directly on the muscle. It is concluded that although 5-HT relaxes the internal anal sphincter, it does not participate in the rectoanal reflex.


Utility of SCORE2 risk algorithm for predicting life course accelerated frailty and physical function decline.
Frailty is a dynamic process that increases with ageing, while it remains unclear whether cardiovascular disease (CVD) risk algorithm could predict life course dynamic frailty trajectories, for example, the longitudinal patterns of how frailty evolves with time. We intended to examine the predictive utility of the Systemic Coronary Risk Estimation 2 (SCORE2) algorithm for life course accelerated frailty and physical function decline, in comparison with the precedent SCORE algorithm. Longitudinal data regarding accumulation of deficits frailty index (FI) and physical function (grip strength, gait speed, peak expiratory flow and timed chair rises) were drawn from the English Longitudinal Study of Ageing (ELSA) and Health and Retirement Study (HRS), two nationally representative cohorts with community-dwelling adults aged ≥50 years. SCORE and SCORE2 were calculated at baselines following European Society of Cardiology guidelines. A group-based trajectory modelling approach was used for identifying potential life course frailty trajectories, based on 14- and 12-year FI data in the ELSA and HRS. Modified Poisson regression and linear mixed model were applied for analysing associations between SCORE2 with accelerated frailty trajectory and physical function decline, respectively. Receiver operating characteristic curve (ROC) analysis was conducted to evaluate predictive utility for accelerated frailty increase trajectory of SCORE and SCORE2, with the area under the curve (AUC) compared using the paired DeLong's test. A total of 4834 participants from the ELSA and 7815 participants from the HRS were included (mean age: 64.0 ± 9.2 and 65.4 ± 9.9 years; men: 44.3% and 41.4%, respectively). Three frailty trajectories were consistently identified in both cohorts: (1) stable frailty increase (n = 3026 in ELSA and 4004 in HRS); (2) moderate frailty increase (n = 1325 in ELSA and 2955 in HRS); (3) accelerated frailty increase (n = 483 in ELSA and 856 in HRS). Each 10% increment in SCORE2 risk was associated with the higher risk of accelerated frailty increase (risk ratio [RR]: 3.58, 95% confidence interval [CI] [3.22, 3.98], P < 0.001 in ELSA; RR: 1.61, 95% CI [1.56, 1.67], P < 0.001 in HRS) and faster declines in all physical function measurements. SCORE2 algorithm showed good accuracy for predicting accelerated frailty increase (area under the curve [AUC] in ELSA: 0.759; HRS: 0.744), with better performance than the SCORE (AUC in ELSA: 0.729; HRS: 0.700) in both cohorts (P < 0.001 for comparison). SCORE2 algorithm could serve good utility for predicting life course accelerated frailty increase and physical function decline among community-dwelling non-frail adults aged ≥50 years.


Enhanced glypican-3 expression differentiates the majority of hepatocellular carcinomas from benign hepatic disorders.
Hepatocellular carcinoma (HCC) is a common malignant tumour worldwide, and its differential diagnosis from benign lesions of the liver is often difficult yet of great clinical importance. In the present study, we analysed whether glypican-3 is useful in differentiating between benign and malignant liver diseases and whether it influences the growth behaviour of HCC. Northern blot analysis and in situ hybridisation. Northern blot analysis indicated that expression of glypican-3 mRNA was either low or absent in normal liver, in focal nodular hyperplasia (FNH), and in liver cirrhosis. In contrast, expression of glypican-3 mRNA was markedly increased in 20 of 30 and moderately increased in five of 30 HCC samples. The average increase in glypican-3 mRNA expression in HCC was significant compared with expression in normal liver (21.7-fold increase, p<0.01). In comparison with FNH or liver cirrhosis, glypican-3 mRNA expression in HCC was increased 7.2- (p<0.05) and 10.8-fold (p<0.01), respectively. In addition, pushing HCCs exhibited significantly higher glypican-3 mRNA expression than invading tumours (p<0.05). In situ hybridisation analysis demonstrated weak expression of glypican-3 mRNA in normal hepatocytes and bile ductular cells, and weak to occasionally moderate signals in hepatocytes forming nodules of liver cirrhosis and in regenerated hepatic nodules of FNH. In contrast, glypican-3 in situ hybridisation signals were intense in hepatic cancer cells with even higher levels in pushing HCCs than in invading HCCs. These findings suggest that glypican-3, in many cases, has the potential to differentiate between benign and malignant liver diseases.


Stretchable Biofuel Cells as Wearable Textile-based Self-Powered Sensors.
Highly stretchable textile-based biofuel cells (BFCs), acting as effective self-powered sensors, have been fabricated using screen-printing of customized stress-enduring inks. Due to synergistic effects of nanomaterial-based engineered inks and the serpentine designs, these printable bioelectronic devices endure severe mechanical deformations, e.g., stretching, indentation, or torsional twisting. Glucose and lactate BFCs with the single enzyme and membrane-free configurations generated the maximum power density of 160 and 250 µW cm-2 with the open circuit voltages of 0.44 and 0.46 V, respectively. The textile-BFCs were able to withstand repeated severe mechanical deformations with minimal impact on its structural integrity, as was indicated from their stable power output after 100 cycles of 100% stretching. By providing power signals proportional to the sweat fuel concentration, these stretchable devices act as highly selective and stable self-powered textile sensors. Applicability to sock-based BFC and self-powered biosensor and mechanically compliant operations was demonstrated on human subjects. These stretchable skin-worn "scavenge-sense-display" devices are expected to contribute to the development of skin-worn energy harvesting systems, advanced non-invasive self-powered sensors and wearable electronics on a stretchable garment.


Are occupational factors important determinants of socioeconomic inequalities in musculoskeletal pain?
The aim of this study was to quantify socioeconomic inequalities in low-back pain, neck-shoulder pain, and arm pain in the general working population in Oslo and to examine the impact of job characteristics on these inequalities. All economically active 30-, 40-, and 45-year-old persons who attended the Oslo health study in 2000-2001 and answered questions on physical job demands, job autonomy, and musculoskeletal pain were included (N=7293). Occupational class was used as an indicator of socioeconomic status. The lower occupational classes were compared with higher grade professionals, and prevalences, prevalence ratios, prevalence differences, and population attributable fractions were calculated. There were marked, stepwise socioeconomic gradients for musculoskeletal pain, steeper for the men than for the women. The relative differences (prevalence ratios) were larger for low-back pain and arm pain than for neck-shoulder pain. The absolute differences (prevalence differences) were the largest for low-back pain. Physical job demands explained a substantial proportion of the absolute occupational class inequalities in low-back pain, while job autonomy was more important in explaining the inequalities in neck-shoulder pain and arm pain. The estimated population attributable fractions supported the impact of job characteristics at the working population level, especially for low-back pain. In this cross-sectional study, physical job demands and job autonomy explained a substantial proportion of occupational class inequalities in self-reported musculoskeletal pain in the working population in Oslo. This finding indicates that the workplace may be an important arena for preventive efforts to reduce socioeconomic inequalities in musculoskeletal pain.


Acoustic radiation force impulse imaging of kidneys - a phantom study.
Since there have been only few works reporting the diagnosis of kidneys using Acoustic Radiation Force Impulse technique and those works do not provide consistent results of shear wave velocity measurements in renal tissue, we have decided to use kidney phantoms with known properties to examine the reliability of the method itself in a controlled setup similar to kidneys examination. Four gelatin-based phantoms imitating different clinical situations were manufactured - two with thick and two with thin renal cortex, each type at a depth similar to a normal-weight or overweight patient. For each phantom, a series of interest points was chosen and for each point 20 Shear Wave Velocity measurements were taken using the build-in Virtual Touch Tissue Quantification™ tool in a Siemens Acuson S2000 ultrasound scanner equipped with a 6C1 HD Transducer (Siemens Mountainview, USA). Mean Shear Wave Velocity values obtained for all the examined points ranged from 2.445 to 3.941 m/s, with standard deviation exceeding 0.1 in only one case out of 29 points, but differing significantly between all points. The obtained results indicate that the method is highly reliable as long as the measurement volume contains a uniform tissue region. If the measurement window covers a region with different properties even partially, the obtained results are affected. The variance of measured values on the other hand is not affected by the said non-uniformity of material under examination. Furthermore, the variance of measured values does not show a clear dependency on the depth at which the shear wave velocities are measured.


Biochemical outcomes and predictive factors by risk group after permanent iodine-125 seed implantation: Prospective cohort study in 2,316 patients.
To evaluate the biochemical freedom from failure (bFFF) by risk group and treatment modality and the predictive factors of bFFF by risk group in patients with prostate cancer undergoing permanent seed implantation (PI) with or without external beam radiation therapy (EBRT) in a nationwide prospective cohort study (Japanese Prostate Cancer Outcome Study of Permanent Iodine-125 [I-125] Seed Implantation) in Japan during the first 2 years. The analyses included 2,316 participants in 42 institutions; bFFF was evaluated using the Phoenix definition and calculated using the Kaplan-Meier method, and the Cox proportional hazards model was used to identify the factors associated with bFFF. Median followup period was 60.0 months. The 5-year bFFF rates in all patients, 1,028 low-risk patients, 1,114 intermediate-risk patients, and 133 high-risk patients were 93.6%, 94.9%, 92.7%, and 91.1%, respectively. The 5-year bFFF rates in the PI group and EBRT combination therapy group were 93.7% and 93.3%, respectively. In a multivariate analysis, younger age, higher Gleason score (GS), higher percent positive biopsies (%PB), and lower prostate V100 (p = 0.0012, 0.0030, 0.0026, and 0.0368) in all patients; younger age, higher pretreatment prostate-specific antigen, and lower prostate V100 (p = 0.0002, 0.0048, and 0.0012) in low-risk patients; higher GS, higher %PB, and no hormonal treatment (p = 0.0005, 0.0120, and 0.0022) in intermediate-risk patients; and higher GS and higher %PB (p = 0.0329 and 0.0120) in high-risk patients were significantly associated with bFFF. PI with or without EBRT resulted in excellent short-term biochemical outcomes in all risk groups, especially in high-risk patients. Age, pretreatment prostate-specific antigen, and prostate V100 in low-risk patients; GS, %PB, and hormonal treatment in intermediate-risk patients; and GS and %PB in high-risk patients were independently affected bFFF.


Method for improved Illumina sequencing library preparation using NuGEN Ovation RNA-Seq System.
In this study, we tested the NuGEN Ovation RNA-Seq System for library preparation followed by next-generation sequencing on an Illumina GAIIx. The cDNA product of the NuGEN kit may have significant amounts of ssDNA with hairpin structures that are generated during the amplification process. These structures interfere with efficient downstream end repair, A-tailing, and adapter ligation, all standard steps in post-amplification sequencing library construction. We were able to increase the efficiency of sequencing library yields 4- to 6-fold or greater by treatment of NuGEN-amplified cDNA product with the single-strand endonuclease S1. These results suggest that this treatment effectively cleaves hairpin structures generated during amplification that are resistant to the standard enzyme cocktails used for the end-repair step.


Particle Formation during Water-Vapor Photolysis.
Micrometer-sized particles have been produced by the photolysis of water vapor at wavelengths between 1500 and 1700 angstroms. Although the composition of the particles was not determined, the rate of particle production is consistent with the efficient conversion of water photolysis products to particulate matter. Such a process may be of importance in planetary atmospheres.


[The use of sigma-delta analog-digital conversion in multichannel electrocardiographs].
With advances in the manufacture of super great integrated circuits, a commercially available element base that realizes the sigma-delta (sigma-delta) analog-to-digital conversion principle in one microcircuit has appeared. The sigma-delta analog-to-digital converters (ADC) have a high resolving power (above 12 bits), they are largely based on the signal digital filtration principles, which may moderate requirements for the analog filtration of signals and use direct-current amplifiers as alternatives to instrument ones. For electrocardiograph operation, the sigma-delta ADC of an AD 7716 (Analog Devices) with 22 bits were used in this study. The use of sigma-delta ADC in the electrocardiographs has been shown to be a rather useful alternative of traditional ADC. Moreover, sigma-delta ADC may be also useful for other systems for collecting and processing biomedical signals which are close in their requirements for a frequency range and a dynamic range, i.e. encephalographic, myographic, rheographic and other input signals.


CT-based measurement of visceral adipose tissue volume as a reliable tool for assessing metabolic risk factors in prediabetes across subtypes.
Visceral adipose tissue (VAT) is a well-established risk factor for the development of diabetes in individuals with prediabetes. We aimed to evaluate various adiposity and metabolic indices, including visceral adipose tissue (VAT) volume measured by CT, in individuals with prediabetes, based on their different subtypes. This retrospective study included 78 prediabetes individuals aged ≥ 20 years whose VAT volumes were evaluated by CT. Individuals were classified into prediabetes subtypes: impaired fasting glucose (IFG), impaired glucose tolerance (IGT), and combined IFG and IGT (C-IFG/IGT). We conducted a comparison of the patients' adiposity indices and their associations with one other, as well as with insulin resistance, based on the different prediabetes subtypes. Individuals with C-IFG/IGT had higher levels of visceral obesity than those with IFG or IGT. VAT volume was more strongly associated with insulin resistance than other adiposity indices, regardless of prediabetes subtypes. Additionally, VAT volume and liver attenuation index showed a significant correlation with the other indices studied across the prediabetes subtypes. CT-based measurement of VAT volume may be a valuable tool for evaluating metabolic risk factors among individuals with prediabetes.


Influence of DNA sequence identity on efficiency of targeted gene replacement.
We have developed a system for analyzing recombination between a DNA fragment released in the nucleus from a single-copy plasmid and a genomic target in order to determine the influence of DNA sequence mismatches on the frequency of gene replacement in Saccharomyces cerevisiae. Mismatching was shown to be a potent barrier to efficient gene replacement, but its effect was considerably ameliorated by the presence of DNA sequences that are identical to the genomic target at one end of a chimeric DNA fragment. Disruption of the mismatch repair gene MSH2 greatly reduces but does not eliminate the barrier to recombination between mismatched DNA fragment and genomic target sequences, indicating that the inhibition of gene replacement with mismatched sequences is at least partially under the control of mismatch repair. We also found that mismatched sequences inhibited recombination between a DNA fragment and the genome only when they were close to the edge of the fragment. Together these data indicate that while mismatches can destabilize the relationship between a DNA fragment and a genomic target sequence, they will only do so if they are likely to be in the heteroduplex formed between the recombining molecules.


Association of Missense Mutation in FOLH1 With Decreased NAAG Levels and Impaired Working Memory Circuitry and Cognition.
Altering the metabotropic glutamate receptor 3 (mGluR3) by pharmacology or genetics is associated with differences in learning and memory in animals and humans. GRM3 (the gene coding for mGluR3) is also genome-wide associated with risk for schizophrenia. The neurotransmitter N-acetyl-aspartyl-glutamate (NAAG) is the selective endogenous agonist of mGluR3, and increasing NAAG may improve cognition. Glutamate carboxypeptidase II (GCPII), coded by the gene folate hydrolase 1 (FOLH1), regulates the amount of NAAG in the synapse. The goal of this study was to determine the relationship between FOLH1, NAAG levels, measures of human cognition, and neural activity associated with cognition. The effects of genetic variation in FOLH1 on mRNA expression in human brain and NAAG levels using 7-T magnetic resonance spectroscopy (MRS) were measured. NAAG levels and FOLH1 genetic variation were correlated with measures of cognition in subjects with psychosis and unaffected subjects. Additionally, FOLH1 genetic variation was correlated with neural activity during working memory, as measured by functional MRI (fMRI). A missense mutation in FOLH1 (rs202676 G allele) was associated with increased FOLH1 mRNA in the dorsolateral prefrontal cortex of brains from unaffected subjects and schizophrenia patients. This FOLH1 variant was associated with decreased NAAG levels in unaffected subjects and patients with psychosis. NAAG levels were positively correlated with visual memory performance. Carriers of the FOLH1 variant associated with lower NAAG levels had lower IQ scores. Carriers of this FOLH1 variant had less efficient cortical activity during working memory. These data show that higher NAAG levels are associated with better cognition, suggesting that increasing NAAG levels through FOLH1/GCPII inhibition may improve cognition. Additionally, NAAG levels measured by MRS and cortical efficiency during working memory measured by fMRI have the potential to be neuroimaging biomarkers for future clinical trials.


Lipids and lipid binding proteins: a perfect match.
Lipids serve a great variety of functions, ranging from structural components of biological membranes to signaling molecules affecting various cellular functions. Several of these functions are related to the unique physico-chemical properties shared by all lipid species, i.e., their hydrophobicity. The latter, however, is accompanied by a poor solubility in an aqueous environment and thus a severe limitation in the transport of lipids in aqueous compartments such as blood plasma and the cellular soluble cytoplasm. Specific proteins which can reversibly and non-covalently associate with lipids, designated as lipid binding proteins or lipid chaperones, greatly enhance the aqueous solubility of lipids and facilitate their transport between tissues and within tissue cells. Importantly, transport of lipids across biological membranes also is facilitated by specific (membrane-associated) lipid binding proteins. Together, these lipid binding proteins determine the bio-availability of their ligands, and thereby markedly influence the subsequent processing, utilization, or signaling effect of lipids. The bio-availability of specific lipid species thus is governed by the presence of specific lipid binding proteins, the affinity of these proteins for distinct lipid species, and the presence of competing ligands (including pharmaceutical compounds). Recent studies suggest that post-translational modifications of lipid binding proteins may have great impact on lipid-protein interactions. As a result, several levels of regulation exist that together determine the bio-availability of lipid species. This short review discusses the significance of lipid binding proteins and their potential application as targets for therapeutic intervention.


Left main coronary artery stenosis treatment with two paclitaxel-eluting stents in a patient with cardiac allograft vasculopathy.
Cardiac transplantation is a well defined therapy for end stage heart failure. After the first year of transplantation, allograft coronary artery disease (ACAD) is the second main cause of death. The ACAD is defined as a diffuse process affecting the entire length of epicardial vessels. Once ACAD has been established, treatments such as coronary angioplasty, coronary stenting, and coronary bypass are performed. We present a case of successful stenting of the left main coronary artery (LMCA) in a patient with ACAD. The patient's medical history was significant for heart transplantation due to ischemic heart failure. Four years after transplantation the patient was admitted again due to sudden worsening of New York Heart Association functional class and extreme fatigue. Coronary angiogram showed a severe stenosis in the proximal segment of the LMCA; we performed stenting with a paclitaxel-eluting stent (PES). Six months after the procedure, the patient had an elective angiogram, where we discovered a new severe occlusion distally to the former stent; a second PES was implanted. Fourteen months after the second stenting, a new elective angiogram was performed without evidence of in-stent restenosis. After a 8-year follow-up since transplantation, the patient is free from dyspnea, angina, and adverse cardiovascular events. Our report suggests the efficacy of PES as ACAD treatment of the unprotected LMCA.


Navigating the tip of the genomic iceberg: Next-generation sequencing for plant systematics.
Just as Sanger sequencing did more than 20 years ago, next-generation sequencing (NGS) is poised to revolutionize plant systematics. By combining multiplexing approaches with NGS throughput, systematists may no longer need to choose between more taxa or more characters. Here we describe a genome skimming (shallow sequencing) approach for plant systematics. Through simulations, we evaluated optimal sequencing depth and performance of single-end and paired-end short read sequences for assembly of nuclear ribosomal DNA (rDNA) and plastomes and addressed the effect of divergence on reference-guided plastome assembly. We also used simulations to identify potential phylogenetic markers from low-copy nuclear loci at different sequencing depths. We demonstrated the utility of genome skimming through phylogenetic analysis of the Sonoran Desert clade (SDC) of Asclepias (Apocynaceae). Paired-end reads performed better than single-end reads. Minimum sequencing depths for high quality rDNA and plastome assemblies were 40× and 30×, respectively. Divergence from the reference significantly affected plastome assembly, but relatively similar references are available for most seed plants. Deeper rDNA sequencing is necessary to characterize intragenomic polymorphism. The low-copy fraction of the nuclear genome was readily surveyed, even at low sequencing depths. Nearly 160000 bp of sequence from three organelles provided evidence of phylogenetic incongruence in the SDC. Adoption of NGS will facilitate progress in plant systematics, as whole plastome and rDNA cistrons, partial mitochondrial genomes, and low-copy nuclear markers can now be efficiently obtained for molecular phylogenetics studies.


Health and education of children with albinism in Zimbabwe.
Albinism is a relatively common genetic condition in Zimbabwe, a tropical country in southern Africa. Those affected have little pigment in their hair, skin or eyes, in sharp contrast to the normal dark pigmentation. This article describes the responses to a self-report questionnaire covering health, social and educational aspects completed by 138 schoolchildren with albinism living in rural areas of Zimbabwe. They reported persistent skin and a myriad of eye problems. Relationships between siblings appeared good, although problems of antagonism, avoidance and fear were encountered among strangers. Knowledge about albinism was patchy; pupils were keen to be better informed. This research highlights the need for widespread dissemination of accurate information about the genetics and health management of albinism to counter the many myths and misconceptions surrounding this condition. A management programme to promote the health and education of these children in southern Africa is proposed. In conclusion, this study indicated that pupils with albinism could participate in mainstream education, with appropriate intervention to help them manage the problems associated with their low vision and sensitive skins.


Molecular cloning, characterization, and downregulation of an acyltransferase that catalyzes the malonylation of flavonoid and naphthol glucosides in tobacco cells.
Tobacco cells (Nicotiana tabacum L. Bright Yellow T-13) exposed to harmful naphthols accumulate them as glucosylated and further modified compounds [Taguchi et al. (2003a) Plant Sci. 164, 231-240]. In this study, we identified the accumulated compounds to be 6'-O-malonylated glucosides of naphthols. Cells treated with various phenolic compounds accumulated the flavonoids mainly as malonylglucosides. To clarify the function of this malonylation in tobacco, we isolated the cDNA encoding a malonyltransferase (NtMaT1) from a cDNA library derived from tobacco cells. The heterologous expression of the gene in Escherichia coli revealed that the recombinant enzyme had malonyltransferase activity against several phenolic glucosides such as flavonoid 7-O-glucosides, flavonoid 3-O-glucosides and naphthol glucosides. The substrate preference of the enzyme was similar to that of the tobacco cell extract. Malonylation activity in the transgenic cells markedly decreased with the suppression of the expression of NtMaT1 mRNA in tobacco BY-2 cells by RNA interference. The compounds administered to the transgenic cells were accumulated in the cells as glucosides or other modified compounds in place of malonylglucosides. These results show that NtMaT1 is the main catalyst of malonylation on glucosides of xenobiotic flavonoids and naphthols in tobacco plants.


Identification of Campylobacter jejuni ATCC 43431-specific genes by whole microbial genome comparisons.
This study describes a novel approach to identify unique genomic DNA sequences from the unsequenced strain C. jejuni ATCC 43431 by comparison with the sequenced strain C. jejuni NCTC 11168. A shotgun DNA microarray was constructed by arraying 9,600 individual DNA fragments from a C. jejuni ATCC 43431 genomic library onto a glass slide. DNA fragments unique to C. jejuni ATCC 43431 were identified by competitive hybridization to the array with genomic DNA of C. jejuni NCTC 11168. The plasmids containing unique DNA fragments were sequenced, allowing the identification of up to 130 complete and incomplete genes. Potential biological roles were assigned to 66% of the unique open reading frames. The mean G+C content of these unique genes (26%) differs significantly from the G+C content of the entire C. jejuni genome (30.6%). This suggests that they may have been acquired through horizontal gene transfer from an organism with a G+C content lower than that of C. jejuni. Because the two C. jejuni strains differ by Penner serotype, a large proportion of the unique ATCC 43431 genes encode proteins involved in lipooligosaccharide and capsular biosynthesis, as expected. Several unique open reading frames encode enzymes which may contribute to genetic variability, i.e., restriction-modification systems and integrases. Interestingly, many of the unique C. jejuni ATCC 43431 genes show identity with a possible pathogenicity island from Helicobacter hepaticus and components of a potential type IV secretion system. In conclusion, this study provides a valuable resource to further investigate Campylobacter diversity and pathogenesis.


Muscarinic receptors on nerves and muscles in opossum esophagus muscularis mucosa.
The muscarinic receptors of muscularis mucosa have some recognition properties that suggest they resemble receptors of the M1 subtype. The nerves of these tissues also contain muscarinic receptors which inhibit tonic contractions caused by release of a substance-P-like material by field stimulation. These receptors also appear to be M1 in type as they are maximally activated by McNeil A343 as well as by carbachol (pD2, 5.5 and 7.5, respectively). They are also inhibited by pirenzepine, as well as by atropine (negative logarithms of the required dose for 50% inhibition or potentiation, 6.6-6.7 compared with 8.2-8.3). Hexahydrosiladifenidol, an antagonist selective or M2 receptors of guinea pig ileum, had a low (approximately 7.1) pA2 value for antagonism of both agonists in smooth muscle in this tissue. However, it was closer to atropine in potency with respect to potentiating tonic responses to field stimulation or to inhibiting phasic responses to field stimulation than it was to antagonizing smooth muscle contractions. Thus, atropine was about 40 times more potent than pirenzepine and 2-5 times more potent than hexahydrosilafenidol. There were some quantitative differences in the effectiveness of these three antagonists in blocking the phasic (acetylcholine-mediated) response to field stimulation. Atropine was 70-100 times more potent than pirenzepine and 8-25 times more potent than hexahydrosiladifenidol. This greater potency difference for inhibition of phasic contractions compared with potentiation of tonic contractions was discussed. This tissue appears to be one of the first smooth muscles in which both nerves and muscles contain muscarinic receptors with some recognition properties resembling those of the M1 subtype.


Dietary supplements for the lactating mother: influence on the trace element content of milk.
Milk production is a complex process where nutritional factors interact with structural hormonal and behavioural influences. In recent years important advances have been made in understanding the role of the nutritional status of lactating women on the outcome of breastfeeding. Many questions remain unanswered about the exact requirement of trace elements for lactating mothers. The effect of dietary zinc, copper and iodine supplements on the milk concentration of these micronutrients was studied. The supplementation trial employed a specific balanced nutritional supplement prepared for the nursing mothers. The study was carried out on women living in Ferrara and its surrounding area. The population under study was healthy Italian mothers, of good socioeconomic status, and their normal infants. In total, 32 women were enrolled in the study and 22 completed it. The infants (9F, 13M) were full-term, healthy singletons and were put to breast within 12 h of birth. All women who finished the study completed a 3 d dietary record. Nutrient analysis revealed the following mean daily dietary trace element intake in the lactating mothers: zinc = 12 mg, copper = 1.4 mg and iodine = 145 microg. The zinc and copper dietary intake was in agreement with the daily intake proposed for nursing Italian mothers, while the daily intake of iodine was below the recommended intake of 200 microg. The breastfeeding mothers were placed in 2 groups, with 7 primiparas and 4 multiparas per group: lactating women eating a traditional Italian diet without vitamin and mineral supplements, and lactating women enrolled in the nutrification programme and given a nutritional supplement to their traditional diet. The supplement (PerMamma Abbott) provided 20mg zinc sulfate, 2mg copper sulfate and 116 microg potassium iodide. These quantities cover about 60-90% of the recommended intake for nursing Italian mothers. Samples of 10 ml of milk were collected at 3, 30, 90d postpartum. Zinc milk concentrations declined significantly over the study period for all lactating subjects, without differences in the rate of decline between the women who started supplementation during lactation and those who did not. Copper did not change during the first month of lactation, then declined at day 90 in supplemented and unsupplemented women, without significant differences between the two groups. An early sharp decline in milk iodine occurred in all lactating subjects, independently of iodine supplementation. After the first month of lactation breast milk iodide levels remained stable in all subjects under study. No significant differences between the two study groups were observed. The lack of correlation between the iodide level in breast milk and maternal dietary intake of iodine is not in agreement with previously published reports. The present results indicate that in healthy, well-nourished lactating Italian women, whose diet is adequate, the levels of zinc, copper and iodine in milk are not influenced by short-term supplementary intakes and that the milk levels of the trace elements studied are maintained over different levels of intake. Further research and examination by longitudinal studies are needed to establish the exact relationship between the amount of iodine furnished to the nursing mother and the iodine content of human milk. The role of compensatory homeostatic mechanisms which act during lactation needs further consideration and closer scrutiny.


A sparsity-controlled vector autoregressive model.
Vector autoregressive (VAR) models constitute a powerful and well studied tool to analyze multivariate time series. Since sparseness, crucial to identify and visualize joint dependencies and relevant causalities, is not expected to happen in the standard VAR model, several sparse variants have been introduced in the literature. However, in some cases it might be of interest to control some dimensions of the sparsity, as e.g. the number of causal features allowed in the prediction. To authors extent none of the existent methods endows the user with full control over the different aspects of the sparsity of the solution. In this article, we propose a versatile sparsity-controlled VAR model which enables a proper visualization of potential causalities while allows the user to control different dimensions of the sparsity if she holds some preferences regarding the sparsity of the outcome. The model coefficients are found as the solution to an optimization problem, solvable by standard numerical optimization routines. The tests performed on both simulated and real-life time series show that our approach may outperform a greedy algorithm and different Lasso approaches in terms of prediction errors and sparsity.